Alkaline Phosphatase Antibody | AbD25296_hIgG2

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Anti BAP specificity ELISA

A microtiter plate was coated overnight with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Bacterial Alkaline Phosphatase:HRP Antibody (clone AbD25296_hIgG2) at a concentration of 2 μg/ml followed by Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.

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Western blot using HSPA8 primary antibody

Western blot analysis was performed by loading 25 μl Hela lysate (lanes 2 and 3) and Precision Plus Protein™ Prestained Standards (lane 1) onto a AnykD™ Criterion™ TGX Stain-Free™ Protein Gel. The gel was run under reducing conditions and proteins were transferred to a nitrocellulose membrane. Lane 2 was probed with Anti-HSPA8 Antibody, clone AbD28028 Fab-A-FH format, at a concentration of 10 μg/ml. Lanes 2 and 3 were then probed with Human Anti-Bacterial Alkaline Phosphatase:HRP (clone AbD25296_hIgG2, HCA275P) at dilution of 1/1000. Visualization was carried out using the ChemiDoc™ MP Imaging System with an auto exposure. The Western blot bands (red) and protein standards (green) have been pseudocolored. Fab-A-FH format is a bivalent bacterial alkaline phosphatase fusion antibody (DYKDDDDK- and His-6-tags).

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Anti BAP specificity titration ELISA

A microtiter plate was coated overnight with recombinant GFP antibody (His-tagged) at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using primary human anti-GFP antibodies in different bivalent Fab formats (e.g. Fab-A-V5Sx2) titrated to the given concentrations in PBST, followed by Human Anti-Bacterial Alkaline Phosphatase:HRP Antibody (clone AbD25296_hIgG2) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of triplicate measurements.

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  • Human anti Bacterial Alkaline Phosphatase:HRP
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    This HRP conjugated Anti-Bacterial Alkaline Phosphatase Antibody is recommended for use as a secondary detection reagent with HuCAL antibodies in Fab-A and Fab-Max formats.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD25296_hIgG2
    • Isotype
      IgG2
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA275PE, WBdatasheet pdfdatasheet pdf0.1 mg
      HCA275P
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
      -
      • Human Anti-Bacterial Alkaline Phosphatase Antibody, clone AbD25296_hIgG2 recognizes bacterial alkaline phosphatase (BAP), a periplasmic enzyme which catalyzes the release of 5'- and 3'-phosphate groups from DNA, RNA and nucleotides. BAP can also dephosphorylate a number of proteins and alkaloids, and has an optimum pH of 8.0. BAP is encoded by the phoA gene, and forms a homodimer of two identical ~47 kDa subunits.

        Fab-A and Fab-Max format antibodies generated using HuCAL® technology are formed via dimerization of BAP. The Anti-BAP Antibody HCA275P can be used as a secondary detection reagent with HuCAL antibodies in Fab-A and Fab-Max formats (Fab-Max is a modified BAP fusion antibody with inactivated enzymatic activity).

        View further information about HuCAL antibody formats.
      • Intended Use
      • Target Species
        Bacterial
      • Product Form
        Human IgG2 antibody (lambda light chain) selected from the HuCAL® phage display library and expressed in a human cell line. Conjugated to horseradish peroxidase (HRP) - liquid
      • Reconstitution
      • Preparation
        Purified IgG prepared by affinity chromatography on Protein A
      • Preservative Stabilisers
        0.01% Thiomersal
        HRP Stabiliser (BUF052A)
      • Immunogen
        Bacterial Alkaline Phosphatase
      • Purity
      • Affinity
        The monovalent intrinsic affinity of this antibody was measured as KD=38pM by real time, label-free molecular interaction analysis on immobilized HuCAL antibody in the Fab-A-FH format
      • Approx. Protein Concentrations
        IgG concentration 1.0 mg/ml
      • Reagents In The Kit
      • Preparing The Antibody
      • Test Principle
      • Buffer Solution
        Phosphate buffered saline
      • Storage
        Store at +4oC. DO NOT FREEZE
        This product should be stored undiluted. This product is photosensitive and should be protected from light. Should this product contain a precipitate we recommend microcentrifugation before use
      • Shelf Life
        12 months from date of despatch
      • GO Terms
        oxidoreductase activity, acting on phosphorus or arsenic in donors
        hydrogenase (acceptor) activity
        alkaline phosphatase activity
        protein binding
        periplasmic space
        zinc ion binding
        magnesium ion binding
      • UniProt
      • Acknowledgements
        Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany.
      • Regulatory
        For research purposes only
      • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

      • Application NameYesNoMin DilutionMax Dilution
        ELISA
        Western Blotting1/1,0001/10,000

      • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
      • Technical Advice
        Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
      • Recommended Protocol
      • ELISA
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Alkaline Phosphatase Antibody Formats

      Formats Clone Applications Sizes available
      Alkaline Phosphatase Antibody : HRP AbD25296_hIgG2 E, WB 0.1 mg
      • Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)

      Recommended Secondary Antibody

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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